1-磷酸鞘氨醇受体2通过AKT/mTOR通路调节线粒体功能参与Aβ_(25-35)诱导的人神经母细胞瘤细胞SH-SY5Y损伤  

S1PR2 Regulates Mitochondrial Function through the AKT/mTOR Pathway to Promote Aβ_(25-35)Damage of SH-SY5Y Cells

作　　者：肖志强 杨柳 黄睿[3] 黄斌[3] 李晓佳[3] 王晓平[3] XIAO Zhi-Qiang;YANG Liu;HUANG Rui;HUANG Bin;LI Xiao-Jia;WANG Xiao-Ping(Department of Neurology,Chengdu University of Traditional Chinese Medicine,Chengdu,610075,China;Department of Neurology,Medical School of University of Electronic Science and Technology of China,Chengdu,610054,China;Department of Neurology,Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital,Chengdu 610072,China)

机构地区：[1]成都中医药大学神经内科,成都610075 [2]电子科技大学医学院神经病学,成都610054 [3]四川省医学科学院四川省人民医院神经内科,成都610072

出　　处：《中国生物化学与分子生物学报》2024年第10期1453-1461,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：四川省自然科学基金(No.2022NSFSC0677);四川省科技厅应用基础研究(No.2020YJ0457);四川省科技厅重点研发项目(No.2023YFS0212)资助。

摘　　要：阿尔兹海默症(Alzheimer’s disease,AD)是一种与年龄相关的认知功能下降的神经退行性疾病。1-磷酸鞘氨醇受体2(sphingosine-1-phosphate receptor 2,S1PR2)参与多种细胞过程,被证实在神经系统发育中发挥重要作用。本文旨在探究S1PR2对Aβ_(25-35)诱导的AD细胞模型损伤的作用及可能机制。研究通过Aβ_(25-35)诱导SH-SY5Y细胞构建细胞损伤模型,并且构建靶向S1PR2的干扰序列用于干预细胞中S1PR2的表达。Western印迹及RT-PCR检测S1PR2蛋白及基因表达,发现Aβ_(25-35)诱导的细胞模型中S1PR2蛋白及基因表达均显著增加(P<0.01),S1PR2干预后模型组内S1PR2蛋白及基因表达显著降低(P<0.001)。CCK8检测细胞增殖活力,流式细胞术检测细胞凋亡,结果显示,S1PR2干预后显著增加Aβ_(25-35)诱导的SH-SY5Y细胞的增殖活性,减少细胞凋亡(P<0.01)。Western印迹检测细胞中APP、Tau、p-Tau和PSD95的表达,结果显示,S1PR2干预后显著降低模型组细胞内APP、Tau和p-Tau的表达,增加突触蛋白PSD95的表达,可显著改善Aβ_(25-35)诱导的细胞损伤(P<0.001)。另外,试剂盒检测细胞中ATP的产生,流式细胞术检测ROS含量及线粒体膜电位以分析细胞的线粒体功能,结果显示,Aβ_(25-35)诱导SH-SY5Y细胞显著降低了细胞中ATP的产生,增加了ROS含量,减少了线粒体膜电位(P<0.001)。S1PR2干预后显著增加Aβ_(25-35)诱导的细胞模型中ATP的产生,降低ROS含量,增加线粒体膜电位(P<0.001)。最后,通过Western印迹检测AKT/mTOR通路蛋白质表达,结果显示,Aβ_(25-35)诱导SH-SY5Y细胞促进了p-AKT/AKT及p-mTOR/mTOR的表达,S1PR2干预后显著抑制AKT/mTOR通路的激活(P<0.001)。总而言之,S1PR2可能通过促进AKT/mTOR通路调节线粒体功能参与Aβ_(25-35)诱导的细胞损伤进程。Alzheimer’s disease(AD)is a neurodegenerative disease with age-related cognitive decline.Sphingosine-1-phosphate receptor 2(S1PR2)is involved in a variety of cellular processes and has been shown to play an important role in nervous system development.This study aimed to investigate the effects and possible mechanism of S1PR2 on Aβ_(25-35)induced cell model damage of AD.In this study,SH-SY5Y cells were induced by Aβ_(25-35)to construct a cell damage model,and the expression of S1PR2 in cells was interfered by targeting sequence.The protein and gene expression levels of S1PR2 were detected by Western blot and RT-PCR.It was found that the expression of S1PR2 was significantly increased at mRNA and protein levels in Aβ_(25-35)-induced SH-SY5Y cell model(P<0.01),and its expression was significantly decreased after S1PR2 intervention(P<0.001).The cell proliferation activity was detected by CCK8,and apoptosis was detected by flow cytometry.The results showed that the proliferation activity of Aβ_(25-35)induced SH-SY5Y cells was significantly increased,and apoptosis was decreased after S1PR2 intervention(P<0.01).The protein levels of APP,Tau,p-Tau,and PSD95 in cells were detected by Western blot to analyze the effect of S1PR2 on the pathology of AD.It was found that after S1PR2 intervention,the expressions of APP,Tau,and p-Tau in the AD cell model were significantly decreased(P<0.001),and the expression of synaptic protein PSD95 was increased(P<0.001),which could significantly improve the pathological damage in Aβ_(25-35)-induced SH-SY5Y cell model.In addition,ATP production was detected by the kit,and ROS content and mitochondrial membrane potential were detected by flow cytometry to analyze the mitochondrial function.Results found that ATP production and mitochondrial membrane potential was significantly decreased,whereas the ROS content was increased in Aβ_(25-35)induced SH-SY5Y cells(P<0.001).Intervention with S1PR2 significantly increased ATP production and mitochondrial membrane potential,but decreased ROS

关 键 词：1-磷酸鞘氨醇受体2 β淀粉样蛋白_(25-35) 阿尔兹海默症 线粒体功能 AKT/mTOR通路 

分 类 号：Q421[生物学—神经生物学]